DOCSLIB.ORG

Balance Between Senescence and Apoptosis Is Regulated by Telomere Damage–Induced Association Between P16 and Caspase-3

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Balance Between Senescence and Apoptosis Is Regulated by Telomere Damage–Induced Association Between P16 and Caspase-3 JBC Papers in Press. Published on May 10, 2018 as Manuscript RA118.003506 The latest version is at http://www.jbc.org/cgi/doi/10.1074/jbc.RA118.003506 Balance between senescence and apoptosis is regulated by telomere damage–induced association between p16 and caspase-3 Shanmugam Panneer Selvam1,2, Braden M. Roth1,2, Rose Nganga1,2, Jisun Kim1,2, Marion A. Cooley3,#, Kristi Helke4, Charles D. Smith5, and Besim Ogretmen1,2* 1Department of Biochemistry and Molecular Biology, 2Hollings Cancer Center, 3Department of Regenerative Medicine, 4Department of Comparative Medicine, Medical University of South Carolina, 86 Jonathan Lucas Street, Charleston, SC 29425; 5Department of Pharmacology, Pennsylvania State University, 500 University Drive, Hershey, PA 17033. #Current Address: Department of Oral Biology, Augusta University, Augusta, GA 30912. *Address correspondence to: [email protected] Running title: p16-caspase-3 complex and senescence Key words: apoptosis; senescence; sphingolipid; sphingosine 1-phosphate; telomere damage Downloaded from Abstract damage–induced apoptosis, indicating that an Telomerase activation protects cells from telomere association between p16 and caspase-3 damage by delaying senescence and inducing cell proteinsforces senescence induction by inhibiting immortalization, whereas telomerase inhibition caspase- 3 activation and apoptosis. These results http://www.jbc.org/ mediates rapid senescence or apoptosis. However, suggest that p16 plays a direct role in telomere the cellular mechanisms that determine telomere damage–dependent senescence by limiting damage–dependent senescence versus apoptosis apoptosis via binding to caspase-3, revealing a induction are largely unknown. Here, we direct link between telomere damage–dependent demonstrate that telomerase instability mediated senescence and apoptosis with regards to aging by guest on May 31, 2018 by silencing of sphingosine kinase 2 (SPHK2) and and cancer. sphingosine 1-phosphate (S1P), which binds and stabilizes telomerase, induces telomere damage– Introduction dependent caspase-3 activation and apoptosis, but Telomerase is composed of a catalytic reverse not senescence, in p16-deficient lung cancer cells transcriptase (TERT) and telomeric template RNA or tumors. These outcomes were prevented by (TR), which plays a key role in the elongation and knockdown of a tumor-suppressor protein, maintenance of telomeres at chromosome ends (1- transcription factor 21 (TCF21), or by ectopic 4). Shelterin complex proteins associate with expression of WT human telomerase reverse telomere repeats for protection from inducing transcriptase (hTERT), but not mutant hTERT DNA damage response to overcome replicative with altered S1P binding. Interestingly, SphK2- end protection problem (5-8). Telomerase is highy deficient mice exhibited accelerated aging and expressed in stem cells, while it is inactivated due telomerase instability that increased telomere to epigenetic silencing of TERT upon damage and senescence via p16 activation differentiation in most somatic cell types (9,10). especially in testes tissues, but not in apoptosis. However, telomerase is reactivated in majority of Moreover, p16 silencing in SphK2-/- mouse cancer cells, inducing cell immortalization and embryonic fibroblasts activated caspase-3 and tumorigenesis (11-14). Recent studies revealed a apoptosis without inducing senescence. Further, clinical relevance of TERT activation in cancer, as ectopic WT p16 expression in p16-deficient A549 mutations within the core promoter region of the lung cancer cells prevented TCF21 and caspase-3 TERT gene provide binding sites for E-twenty-six activation, and resulted in senescence in response (ETS) transcription factors, leading to cancer- to SphK2/S1P inhibition and telomere damage. specific telomerase reactivation (15-17). Mechanistically, a p16 mutant with impaired Activation of telomerase protects cells caspase-3 association did not prevent telomere from telomere damage, delaying senescence and aging process (18,19), whereas targeting (Fig. 1A-B). Similarly, inhibition of SPHK2 using telomerase induces tumor suppressor responses ABC294640 (100 mg/kg, 7 days/week treatment such as senescence or apoptosis (20,21). However, for 21 days) suppressed A549-xenograft-tumor signaling mechanisms that distinctly induce growth compared to vehicle-treated controls (Fig. senescence versus apoptosis in non-cancerous 1A). Interestingly, ABC294640 treatment had no versus cancerous cells in response to telomerase additional effect on A549-xenograft-tumor growth inactivation and telomere damage induction are suppression in response to shRNA-mediated largely unknown. SPHK2 knockdown, supporting its role in SPHK2 Sphingosine 1-phosphate (S1P) is a inhibition for tumor suppression (Fig. 1A). bioactive sphingolipid, generated by sphingosine Suppression of A549-xenograft-tumor growth in kinase 1 or 2 (SPHK1 or SPHK2), that exerts pro- response to SPHK2 knockdown or inhibition was oncogenic signaling via engaging with its G- consistent with decreased hTERT protein protein coupled receptors (S1PR1-5) in an abundance and apoptosis measured by IHC using autocrine or paracrine manner, and/or via directly anti-hTERT antibody and TUNEL assay, binding to regulate its intracellular target proteins respectively, compared to control tumor tissues without receptor signaling (22-25). Intracellular (Fig. 1C-D). In addition, SPHK2 inhibition or targets of SPHK2-generated nuclear S1P include knockdown resulted in increased telomere damage HDAC1/2 (26) and TERT (27). Our previous in A549-xenograft-tumors compared to controls, Downloaded from study described a role for SPHK2-generated S1P measured ex vivo by immunofluorescence using in the stabilization of telomerase via directly anti-TRF2 and anti-g-H2AX antibodies in TIF binding to hTERT involving its D684 residue (27). (telomere damage-induced foci) assay (31) (Fig. These studies also suggested that S1P binding 1E). Moreover, there was no significant difference http://www.jbc.org/ mimicked the phosphorylation of hTERT at Ser921, in the staining of senescence associated-beta which protected hTERT from ubiquitination and galactosidase in shSPHK2 tumors compared to proteasomal degradation (27). However, roles and shScr controls (Fig. 1F). Similar data were also mechanisms by which targeting SPHK2/S1P and obtained when A549-luciferase derived tumors TERT induces senescence versus apoptosis with were grown orthotopically in the lungs of SCID by guest on May 31, 2018 respect to telomere damage signaling and tumor mice after tail vein injection, and inhibition of suppression are largely unknown. Thus, in this SPHK2 using ABC294640 decreased lung tumor study, we have outlined experiments to uncover growth (~95%) and hTERT abundance compared signaling mechanisms that determine responses for to vehicle-treated controls (Supplemental Fig. the induction of senescence versus apoptosis via S1B-D). telomere damage-mediated cellular stress. To assess the clinical relevance of hTERT regulation by SPHK2/S1P signaling, we then Results determined SPHK2 abundance with regard to Inhibition of SPHK2/S1P attenuates hTERT hTERT expression by IHC using commercially abundance and tumor growth available tumor microarrays (TMAs) containing To determine the roles of SPHK2/S1P in lung tumor tissues obtained from patients with the regulation of tumor suppression via controlling NSCLC (n=48) and their adjacent pathologically telomerase and telomere damage, we measured the non-cancerous lung tissues (n=48). The data effects of silencing and/or pharmacologic showed that SPHK2 is significantly over- inhibition of SPHK2/S1P signaling using shRNA- expressed (~2.5-fold) in the majority of NSCLC dependent knockdown or small molecule inhibitor tumors compared to their controls (46/48 tumors, of SPHK2, ABC294640 (28-30) (Supplemental p<0.05) (Supplemental Fig. S2A-B). The data also Fig. S1A) on the growth of A549 xenograft- showed that hTERT was up-regulated (~2.8-fold) derived tumors in SCID mice. ShRNA-mediated in the majority of NSCLC tumors (47/48, p<0.05) knockdown of SPHK2 (~75% mRNA reduction (Supplemental Fig. S2B). Importantly, the compared to Scr-shRNA-expressing tumors, Spearman correlation analysis showed that there is measured by qPCR) almost completely prevented a significant (n=48, p<0.001) association between A549-xenograft-tumor growth compared to Scr- SPHK2 and hTERT expression in these tissues shRNA/A549-xenograft controls after 21 days (Supplemental Fig. S2C). Similar correlation 2 between SPHK2 and hTERT was detected in the activation in response to ABC294640, catalytically majority of tumor tissues obtained from patients inactive mutant SPHK2G212E expression (34) was with small cell lung cancer (SCLC) (Supplemental not protective compared to controls (Fig. 2D-E). Fig. S2D). Thus, these data suggest that targeting Exogenous addition of S1P (5µM, 1h) also clinically relevant and upregulated SPHK2/S1P in prevented ABC294640-mediated caspase 3 tumors inhibits hTERT abundance, which is activation in A549 cells compared to vehicle- associated with increased telomere damage and treated controls (Fig. 2F). apoptosis, leading to tumor growth suppression in Increased telomere damage in response to vivo. genetic loss of SphK2 was also detected in SphK2- /- compared to WT-MEFs (Fig. 3A). Moreover, Targeting SPHK2/S1P signaling mediates shRNA-mediated SPHK2 knockdown resulted in telomere damage and subsequent caspase 3 telomere damage in A549 cells compared to Scr- activation
Load more

Recommended publications
  • Gene Symbol Gene Description ACVR1B Activin a Receptor, Type IB
    Table S1. Kinase clones included in human kinase cDNA library for yeast two-hybrid screening Gene Symbol Gene Description ACVR1B activin A receptor, type IB ADCK2 aarF domain containing kinase 2 ADCK4 aarF domain containing kinase 4 AGK multiple substrate lipid kinase;MULK AK1 adenylate kinase 1 AK3 adenylate kinase 3 like 1 AK3L1 adenylate kinase 3 ALDH18A1 aldehyde dehydrogenase 18 family, member A1;ALDH18A1 ALK anaplastic lymphoma kinase (Ki-1) ALPK1 alpha-kinase 1 ALPK2 alpha-kinase 2 AMHR2 anti-Mullerian hormone receptor, type II ARAF v-raf murine sarcoma 3611 viral oncogene homolog 1 ARSG arylsulfatase G;ARSG AURKB aurora kinase B AURKC aurora kinase C BCKDK branched chain alpha-ketoacid dehydrogenase kinase BMPR1A bone morphogenetic protein receptor, type IA BMPR2 bone morphogenetic protein receptor, type II (serine/threonine kinase) BRAF v-raf murine sarcoma viral oncogene homolog B1 BRD3 bromodomain containing 3 BRD4 bromodomain containing 4 BTK Bruton agammaglobulinemia tyrosine kinase BUB1 BUB1 budding uninhibited by benzimidazoles 1 homolog (yeast) BUB1B BUB1 budding uninhibited by benzimidazoles 1 homolog beta (yeast) C9orf98 chromosome 9 open reading frame 98;C9orf98 CABC1 chaperone, ABC1 activity of bc1 complex like (S. pombe) CALM1 calmodulin 1 (phosphorylase kinase, delta) CALM2 calmodulin 2 (phosphorylase kinase, delta) CALM3 calmodulin 3 (phosphorylase kinase, delta) CAMK1 calcium/calmodulin-dependent protein kinase I CAMK2A calcium/calmodulin-dependent protein kinase (CaM kinase) II alpha CAMK2B calcium/calmodulin-dependent
    [Show full text]
  • A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus
    Page 1 of 781 Diabetes A Computational Approach for Defining a Signature of β-Cell Golgi Stress in Diabetes Mellitus Robert N. Bone1,6,7, Olufunmilola Oyebamiji2, Sayali Talware2, Sharmila Selvaraj2, Preethi Krishnan3,6, Farooq Syed1,6,7, Huanmei Wu2, Carmella Evans-Molina 1,3,4,5,6,7,8* Departments of 1Pediatrics, 3Medicine, 4Anatomy, Cell Biology & Physiology, 5Biochemistry & Molecular Biology, the 6Center for Diabetes & Metabolic Diseases, and the 7Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202; 2Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, Indianapolis, IN, 46202; 8Roudebush VA Medical Center, Indianapolis, IN 46202. *Corresponding Author(s): Carmella Evans-Molina, MD, PhD ([email protected]) Indiana University School of Medicine, 635 Barnhill Drive, MS 2031A, Indianapolis, IN 46202, Telephone: (317) 274-4145, Fax (317) 274-4107 Running Title: Golgi Stress Response in Diabetes Word Count: 4358 Number of Figures: 6 Keywords: Golgi apparatus stress, Islets, β cell, Type 1 diabetes, Type 2 diabetes 1 Diabetes Publish Ahead of Print, published online August 20, 2020 Diabetes Page 2 of 781 ABSTRACT The Golgi apparatus (GA) is an important site of insulin processing and granule maturation, but whether GA organelle dysfunction and GA stress are present in the diabetic β-cell has not been tested. We utilized an informatics-based approach to develop a transcriptional signature of β-cell GA stress using existing RNA sequencing and microarray datasets generated using human islets from donors with diabetes and islets where type 1(T1D) and type 2 diabetes (T2D) had been modeled ex vivo. To narrow our results to GA-specific genes, we applied a filter set of 1,030 genes accepted as GA associated.
    [Show full text]
  • 4-6 Weeks Old Female C57BL/6 Mice Obtained from Jackson Labs Were Used for Cell Isolation
    Methods Mice: 4-6 weeks old female C57BL/6 mice obtained from Jackson labs were used for cell isolation. Female Foxp3-IRES-GFP reporter mice (1), backcrossed to B6/C57 background for 10 generations, were used for the isolation of naïve CD4 and naïve CD8 cells for the RNAseq experiments. The mice were housed in pathogen-free animal facility in the La Jolla Institute for Allergy and Immunology and were used according to protocols approved by the Institutional Animal Care and use Committee. Preparation of cells: Subsets of thymocytes were isolated by cell sorting as previously described (2), after cell surface staining using CD4 (GK1.5), CD8 (53-6.7), CD3ε (145- 2C11), CD24 (M1/69) (all from Biolegend). DP cells: CD4+CD8 int/hi; CD4 SP cells: CD4CD3 hi, CD24 int/lo; CD8 SP cells: CD8 int/hi CD4 CD3 hi, CD24 int/lo (Fig S2). Peripheral subsets were isolated after pooling spleen and lymph nodes. T cells were enriched by negative isolation using Dynabeads (Dynabeads untouched mouse T cells, 11413D, Invitrogen). After surface staining for CD4 (GK1.5), CD8 (53-6.7), CD62L (MEL-14), CD25 (PC61) and CD44 (IM7), naïve CD4+CD62L hiCD25-CD44lo and naïve CD8+CD62L hiCD25-CD44lo were obtained by sorting (BD FACS Aria). Additionally, for the RNAseq experiments, CD4 and CD8 naïve cells were isolated by sorting T cells from the Foxp3- IRES-GFP mice: CD4+CD62LhiCD25–CD44lo GFP(FOXP3)– and CD8+CD62LhiCD25– CD44lo GFP(FOXP3)– (antibodies were from Biolegend). In some cases, naïve CD4 cells were cultured in vitro under Th1 or Th2 polarizing conditions (3, 4).
    [Show full text]
  • Supplementary Table 1. in Vitro Side Effect Profiling Study for LDN/OSU-0212320. Neurotransmitter Related Steroids
    Supplementary Table 1. In vitro side effect profiling study for LDN/OSU-0212320. Percent Inhibition Receptor 10 µM Neurotransmitter Related Adenosine, Non-selective 7.29% Adrenergic, Alpha 1, Non-selective 24.98% Adrenergic, Alpha 2, Non-selective 27.18% Adrenergic, Beta, Non-selective -20.94% Dopamine Transporter 8.69% Dopamine, D1 (h) 8.48% Dopamine, D2s (h) 4.06% GABA A, Agonist Site -16.15% GABA A, BDZ, alpha 1 site 12.73% GABA-B 13.60% Glutamate, AMPA Site (Ionotropic) 12.06% Glutamate, Kainate Site (Ionotropic) -1.03% Glutamate, NMDA Agonist Site (Ionotropic) 0.12% Glutamate, NMDA, Glycine (Stry-insens Site) 9.84% (Ionotropic) Glycine, Strychnine-sensitive 0.99% Histamine, H1 -5.54% Histamine, H2 16.54% Histamine, H3 4.80% Melatonin, Non-selective -5.54% Muscarinic, M1 (hr) -1.88% Muscarinic, M2 (h) 0.82% Muscarinic, Non-selective, Central 29.04% Muscarinic, Non-selective, Peripheral 0.29% Nicotinic, Neuronal (-BnTx insensitive) 7.85% Norepinephrine Transporter 2.87% Opioid, Non-selective -0.09% Opioid, Orphanin, ORL1 (h) 11.55% Serotonin Transporter -3.02% Serotonin, Non-selective 26.33% Sigma, Non-Selective 10.19% Steroids Estrogen 11.16% 1 Percent Inhibition Receptor 10 µM Testosterone (cytosolic) (h) 12.50% Ion Channels Calcium Channel, Type L (Dihydropyridine Site) 43.18% Calcium Channel, Type N 4.15% Potassium Channel, ATP-Sensitive -4.05% Potassium Channel, Ca2+ Act., VI 17.80% Potassium Channel, I(Kr) (hERG) (h) -6.44% Sodium, Site 2 -0.39% Second Messengers Nitric Oxide, NOS (Neuronal-Binding) -17.09% Prostaglandins Leukotriene,
    [Show full text]
  • Prognostic Roles of the Expression of Sphingosine-1-Phosphate Metabolism Enzymes in Non-Small Cell Lung Cancer
    681 Original Article Prognostic roles of the expression of sphingosine-1-phosphate metabolism enzymes in non-small cell lung cancer Yingqin Wang1#, Yaxing Shen2#, Xia Sun3,4, Tinah L. Hong5, Long Shuang Huang6, Ming Zhong1 1Department of Critical Care Medicine, 2Department of Thoracic Surgery, Zhongshan Hospital, Fudan University, Shanghai 200032, China; 3Department of Nephrology, Xuzhou Municipal Hospital Affiliated to Xuzhou Medical University, Xuzhou 221000, China; 4Cancer Institute, Xuzhou Medical University, Xuzhou 221002, China; 5New Trier High School, Winnetka, IL, USA; 6Department of Pharmacology, Shanghai Hospital of Traditional Chinese Medicine, Shanghai 200032, China Contributions: (I) Conception and design: M Zhong, LS Huang; (II) Administrative support: M Zhong; (III) Provision of study materials or patients: LS Huang, Y Wang; (IV) Collection and assembly of data: Y Wang, Y Shen, X Sun; (V) Data analysis and interpretation: Y Wang, Y Shen, TL Hong, X Sun; (VI) Manuscript writing: All authors; (VII) Final approval of manuscript: All authors. #These authors contributed equally to this work. Correspondence to: Ming Zhong, MD, PhD. Department of Critical Care Medicine, Zhongshan Hospital, Fudan University, 180 Fenglin Road, Shanghai 200032, China. Email: [email protected]; Long Shuang Huang, PhD. Department of Pharmacology, Shanghai Hospital of Traditional Chinese Medicine, 274 Zhijiang Middle Rd, Zhabei Qu, Shanghai 200072, China. Email: [email protected]. Background: Sphingosine-1-phosphate (S1P), a bioactive lipid, is generally increased in human non- small cell lung cancer (NSCLC). Evidence has shown that the levels of enzymes in S1P metabolism were associated with clinical outcomes in patients with NSCLC. Nevertheless, the roles of mRNA expression of major enzymes (SPHK1, SPHK2 and SGPL1) in S1P metabolism for predicting outcomes in NSCLC patients have not been determined.
    [Show full text]
  • Whole Egg Consumption Increases Gene Expression Within the Glutathione Pathway in the Liver of Zucker Diabetic Fatty Rats
    Food Science and Human Nutrition Publications Food Science and Human Nutrition 11-3-2020 Whole egg consumption increases gene expression within the glutathione pathway in the liver of Zucker Diabetic Fatty rats Joe L. Webb Iowa State University Amanda E. Bries Iowa State University, [email protected] Brooke Vogel Iowa State University Claudia Carrillo Iowa State University, [email protected] Lily Harvison Iowa State University, [email protected] See next page for additional authors Follow this and additional works at: https://lib.dr.iastate.edu/fshn_hs_pubs Part of the Dietetics and Clinical Nutrition Commons, Endocrinology, Diabetes, and Metabolism Commons, Exercise Science Commons, Food Chemistry Commons, Human and Clinical Nutrition Commons, and the Molecular, Genetic, and Biochemical Nutrition Commons The complete bibliographic information for this item can be found at https://lib.dr.iastate.edu/ fshn_hs_pubs/38. For information on how to cite this item, please visit http://lib.dr.iastate.edu/ howtocite.html. This Article is brought to you for free and open access by the Food Science and Human Nutrition at Iowa State University Digital Repository. It has been accepted for inclusion in Food Science and Human Nutrition Publications by an authorized administrator of Iowa State University Digital Repository. For more information, please contact [email protected]. Whole egg consumption increases gene expression within the glutathione pathway in the liver of Zucker Diabetic Fatty rats Abstract Nutrigenomic evidence supports the idea that Type 2 Diabetes Mellitus (T2DM) arises due to the interactions between the transcriptome, individual genetic profiles, lifestyle, and diet. Since eggs are a nutrient dense food containing bioactive ingredients that modify gene expression, our goal was to examine the role of whole egg consumption on the transcriptome during T2DM.
    [Show full text]
  • Targeting the Sphingosine Kinase/Sphingosine-1-Phosphate Signaling Axis in Drug Discovery for Cancer Therapy
    cancers Review Targeting the Sphingosine Kinase/Sphingosine-1-Phosphate Signaling Axis in Drug Discovery for Cancer Therapy Preeti Gupta 1, Aaliya Taiyab 1 , Afzal Hussain 2, Mohamed F. Alajmi 2, Asimul Islam 1 and Md. Imtaiyaz Hassan 1,* 1 Centre for Interdisciplinary Research in Basic Sciences, Jamia Millia Islamia, Jamia Nagar, New Delhi 110025, India; [email protected] (P.G.); [email protected] (A.T.); [email protected] (A.I.) 2 Department of Pharmacognosy, College of Pharmacy, King Saud University, Riyadh 11451, Saudi Arabia; afi[email protected] (A.H.); [email protected] (M.F.A.) * Correspondence: [email protected] Simple Summary: Cancer is the prime cause of death globally. The altered stimulation of signaling pathways controlled by human kinases has often been observed in various human malignancies. The over-expression of SphK1 (a lipid kinase) and its metabolite S1P have been observed in various types of cancer and metabolic disorders, making it a potential therapeutic target. Here, we discuss the sphingolipid metabolism along with the critical enzymes involved in the pathway. The review provides comprehensive details of SphK isoforms, including their functional role, activation, and involvement in various human malignancies. An overview of different SphK inhibitors at different phases of clinical trials and can potentially be utilized as cancer therapeutics has also been reviewed. Citation: Gupta, P.; Taiyab, A.; Hussain, A.; Alajmi, M.F.; Islam, A.; Abstract: Sphingolipid metabolites have emerged as critical players in the regulation of various Hassan, M..I. Targeting the Sphingosine Kinase/Sphingosine- physiological processes. Ceramide and sphingosine induce cell growth arrest and apoptosis, whereas 1-Phosphate Signaling Axis in Drug sphingosine-1-phosphate (S1P) promotes cell proliferation and survival.
    [Show full text]
  • The Effects of Bone Marrow Adipocytes on Metabolic Regulation in Metastatic Prostate Cancer" (2017)
    Wayne State University Wayne State University Dissertations 1-1-2017 The ffecE ts Of Bone Marrow Adipocytes On Metabolic Regulation In Metastatic Prostate Cancer Jonathan Diedrich Wayne State University, Follow this and additional works at: https://digitalcommons.wayne.edu/oa_dissertations Part of the Oncology Commons Recommended Citation Diedrich, Jonathan, "The Effects Of Bone Marrow Adipocytes On Metabolic Regulation In Metastatic Prostate Cancer" (2017). Wayne State University Dissertations. 1797. https://digitalcommons.wayne.edu/oa_dissertations/1797 This Open Access Dissertation is brought to you for free and open access by DigitalCommons@WayneState. It has been accepted for inclusion in Wayne State University Dissertations by an authorized administrator of DigitalCommons@WayneState. THE EFFECTS OF BONE MARROW ADIPOCYTES ON METASTATIC PROSTATE CANCER CELL METABOLISM AND SIGNALLING by JONATHAN DRISCOLL DIEDRICH DISSERTATION Submitted to the Graduate School of Wayne State University, Detroit, Michigan in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY 2017 MAJOR: CANCER BIOLOGY Approved By: Advisor Date © COPYRIGHT BY JONATHAN DIEDRICH 2017 All Rights Reserved DEDICATION To my Family, Friends, and Wally ii ACKNOWLEDGMENTS When I joined the Podgorski laboratory in April of 2014, I had finished my rotations and spent some time in a collaborating laboratory honing my technical and creative thinking skills to become a valuable asset to her team; however, I was still unprepared for the exciting journey it would be through Izabela’s laboratory over the last three years. I was extremely lucky to have landed in Dr. Podgorski’s laboratory and will be forever thankful for the tremendous support she has given me to aid in my development as an independent investigator.
    [Show full text]
  • Specific NEMO Mutations Impair CD40-Mediated C-Rel Activation and B Cell Terminal Differentiation Ashish Jain,1 Chi A
    Research article Specific NEMO mutations impair CD40-mediated c-Rel activation and B cell terminal differentiation Ashish Jain,1 Chi A. Ma,1 Eduardo Lopez-Granados,1 Gary Means,2 William Brady,2 Jordan S. Orange,3 Shuying Liu,1 Steven Holland,1 and Jonathan M.J. Derry2 1Laboratory of Host Defense, National Institute of Allergy and Infectious Diseases (NIAID), NIH, Bethesda, Maryland, USA. 2Amgen Inc., Seattle, Washington, USA. 3Division of Immunology, Children’s Hospital Boston, Boston, Massachusetts, USA. Hypomorphic mutations in the zinc finger domain of NF-κB essential modulator (NEMO) cause X-linked hyper-IgM syndrome with ectodermal dysplasia (XHM-ED). Here we report that patient B cells are character- ized by an absence of Ig somatic hypermutation (SHM) and defective class switch recombination (CSR) despite normal induction of activation-induced cytidine deaminase (AID) and Iε-Cε transcripts. This indicates that AID expression alone is insufficient to support neutralizing antibody responses. Furthermore, we show that patient B cells stimulated with CD40 ligand are impaired in both p65 and c-Rel activation, and whereas addi- tion of IL-4 can enhance p65 activity, c-Rel activity remains deficient. This suggests that these NF-κB compo- nents have different activation requirements and that IL-4 can augment some but not all NEMO-dependent NF-κB signaling. Finally, using microarray analysis of patient B cells we identified downstream effects of impaired NF-κB activation and candidate factors that may be necessary for CSR and SHM in B cells. Introduction target genes. As a result of its central position in NF-κB signal- Higher vertebrates rely on the diversity of the antibody repertoire ing, large genomic rearrangements in NEMO cause incontinentia to combat infectious pathogens.
    [Show full text]
  • Sphingosine Kinase 1–Mediated Inhibition of Fas Death Signaling in Rheumatoid Arthritis B Lymphoblastoid Cells
    ARTHRITIS & RHEUMATISM Vol. 54, No. 3, March 2006, pp 754–764 DOI 10.1002/art.21635 © 2006, American College of Rheumatology Sphingosine Kinase 1–Mediated Inhibition of Fas Death Signaling in Rheumatoid Arthritis B Lymphoblastoid Cells Xiujun Pi, Shi-Yu Tan, Michael Hayes, Liqun Xiao, James A. Shayman, Song Ling, and Joseph Holoshitz Objective. It is becoming increasingly apparent increased levels of S1P. Real-time PCR analysis showed that B cells play an important role in the pathogenesis higher SPHK-1 mRNA expression levels in RA patients of rheumatoid arthritis (RA). Due to the scarcity of B compared with paired controls. Increased SPHK-1 (but cells in RA, it has been technically difficult to function- not SPHK-2) mRNA levels were observed in synovial ally characterize B cell apoptosis in this disease. As a tissue from RA patients. Competitive inhibitors of necessary first step to identify candidate aberrations, we SPHK reversed the resistance of RA LCLs to Fas- investigated Fas-mediated signaling events in immortal- induced apoptosis. Additionally, resistance to Fas- ized peripheral blood B lymphoblastoid cell lines mediated signaling was reversed by siRNA oligonucleo- (LCLs) from patients with RA and controls. tides specific for SPHK-1 but not by oligonucleotides Methods. Cell death was determined by the MTS specific for SPHK-2. assay, and apoptosis was detected by the TUNEL assay Conclusion. These findings demonstrate disease- and DNA laddering. Proteolytic activation of caspase 3 specific resistance to Fas-mediated death signaling in was determined by immunoblotting, and its enzymatic patients with RA and implicate increased SPHK-1 ac- activity was determined by a fluorometric technique.
    [Show full text]
  • Telomere Maintenance Pathway Activity Analysis Enables Tissue- and Gene-Level Inferences
    bioRxiv preprint doi: https://doi.org/10.1101/2021.02.01.429081; this version posted February 2, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Telomere maintenance pathway activity analysis enables tissue- and gene-level inferences Lilit Nersisyan1,2*, Arman Simonyan1, Hans Binder3, Arsen Arakelyan1,2 1 Bioinformatics Group, Institute of Molecular Biology, National Academy of Sciences, Yerevan, Armenia 2 Pathverse, LLC, Yerevan, Armenia 3 Interdisciplinary Center for Bioinformatics, University of Leipzig, Leipzig, Germany * Correspondence: Lilit Nersisyan Keywords: telomere maintenance mechanisms, telomerase, alternative lengthening of telomeres, pathway signal flow, testis ABSTRACT Telomere maintenance is one of the mechanisms ensuring indefinite divisions of cancer and stem cells. Good understanding of telomere maintenance mechanisms (TMM) is important for studying cancers and designing therapies. However, molecular factors triggering selective activation of either the telomerase dependent (TEL) or the alternative lengthening of telomeres (ALT) pathway are poorly understood. In addition, more accurate and easy-to-use methodologies are required for TMM phenotyping. In this study, we have performed literature based reconstruction of signaling pathways for the ALT and TEL TMMs. Gene expression data were used for computational assessment of TMM pathway activities and compared with experimental assays for TEL and ALT. Explicit consideration of pathway topology makes bioinformatics analysis more informative compared to computational methods based on simple summary measures of gene expression. Application to healthy human tissues showed high ALT and TEL pathway activities in testis, and identified genes and pathways that may trigger TMM activation.
    [Show full text]
  • Sphingolipids and Mitochondrial Dynamic
    cells Review Sphingolipids and Mitochondrial Dynamic Lais Brigliadori Fugio, Fernanda B. Coeli-Lacchini and Andréia Machado Leopoldino * Department of Clinical Analyses Toxicology and Sciences, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Av. do Café s/n, 14040-903 Ribeirão Preto, SP, Brazil; [email protected] (L.B.F.); [email protected] (F.B.C.-L.) * Correspondence: [email protected] Received: 1 December 2019; Accepted: 27 February 2020; Published: 1 March 2020 Abstract: For decades, sphingolipids have been related to several biological functions such as immune system regulation, cell survival, and proliferation. Recently, it has been reported that sphingolipids could be biomarkers in cancer and in other human disorders such as metabolic diseases. This is evidenced by the biological complexity of the sphingolipids associated with cell type-specific signaling and diverse sphingolipids molecules. As mitochondria dynamics have serious implications in homeostasis, in the present review, we focused on the relationship between sphingolipids, mainly ceramides and sphingosine-1-phosphate, and mitochondrial dynamics directed by fission, fusion, and mitophagy. There is evidence that the balances of ceramides (C18 and C16) and S1P, as well as the location of specific ceramide synthases in mitochondria, have roles in mitophagy and fission with an impact on cell fate and metabolism. However, signaling pathways controlling the sphingolipids metabolism and their location in mitochondria need to be better understood in order to propose new interventions and therapeutic strategies. Keywords: ceramide; sphingosine; mitophagy; fission; fusion; mitochondria 1. Introduction Sphingolipids are a class of complex lipids with several functions, including membrane structure and intracellular and extracellular signaling, associated with a large number of processes.
    [Show full text]